An Investigation into the Phytochemical Content and Antioxidant, Antidiabetic, and Wound-Healing Activities of Curculigo latifolia Found in Brunei Darussalam

This present study aimed to investigate the phytochemical content and antioxidant and antidiabetic activities of Curculigo latifolia leaves (CL) and C. latifolia roots (CR) found in Brunei Darussalam. Phytochemical screening showed that CL and CR extracts contain saponins, tannins, glycosides, and terpenoids. CR showed higher total phenolic content (TPC), but lower total flavonoid content (TFC) when compared to CL. The high TPC in CR contributed to its potent radical scavenging activity (RSA) against 2,2-diphenyl-1-picrylhydrazyl (DPPH) radicals and strong ferric reducing antioxidant power (FRAP). Additionally, CR exerted significant inhibition of ∝-glucosidase and ∝-amylase, suggesting a potential link between the chemical compounds and its antioxidant and antidiabetic effects. In the animal study of antihyperglycemic activity, treatment with 250 mg/kg body weight (b.w.) of the CL extract normalised the blood glucose levels and improved body weight gain of alloxan-induced diabetic rats within 14 weeks. Furthermore, our investigation into the wound-healing effects of young C. latifolia leaves (YCL) and matured C. latifolia leaves (MCL) showed a significant reduction in wound size on Day 3, 5, and 7 of the experimental study, indicating its wound-healing potential. Based on our findings, C. latifolia can be consumed as part of a balanced diet due to its antioxidant and antidiabetic properties.


Introduction
Diabetes is a serious chronic medical condition that occurs when a body experiences unregulated blood glucose levels [1].Tis disease is usually associated with defciency in insulin secretion, insulin action, or both [1].Tere are two main types of diabetes, which are type 1 diabetes and type 2 diabetes.Patients with type 1 diabetes usually sufer from absolute defciency of insulin that results from an autoimmune destruction of pancreatic β-cells [2].Meanwhile, type 2 diabetes occurs when there is insulin resistance or when the pancreas does not produce enough insulin [2].Te prevalence of diabetes is increasing over the years, and it poses as a signifcant risk with various complications such as cardiovascular disease, kidney failure, nerve damage, and vision problems [3].Chronic wound (foot ulcers) due to impaired healing is another microvascular complication in diabetes, resulting from physiological processes encompassing vascular, neuropathic, immune, and biochemical elements [4].Additionally, poor circulation of blood, uncontrolled hyperglycemia, and prolonged infammation cause a signifcant delay in the wound-healing process, subsequently increasing the risk of infection, tissue necrosis, gangrene, and nontraumatic amputation [5].Te management of diabetic foot ulcers poses a medical and clinical challenge, and therefore, advancements in wound care technology and diabetes treatment are essential for an enhanced quality of life for afected individuals.
Medicinal plants have gained recognition for their therapeutic properties such as antidiabetic, antiinfammatory, wound-healing, and antimicrobial activities [6].Brunei Darussalam has rich biodiversity and a wide range of fora and fauna, many of which have been often utilised in traditional medicine [7].Despite their longstanding traditional use, scientifc evidence supporting their therapeutic efcacies is still limited.One of the local plants that has been recognised for its healing attributes is Curculigo latifolia, or locally known as lemba, which belongs to the Hypoxidaceae family and genus Curculigo [8].Te plant is commonly found in the Borneo Island and Southeast Asia regions and is traditionally used to treat headaches, mouth thrush, and diarrhoeas [9].A previous study found the root extract to contain various bioactive compounds, such as phloridzin, pomiferin, and cinnamic acid, which exhibited antidiabetic activities through reducing blood glucose levels and increasing insulin secretion [10].Tere have also been reports that revealed that the fruit, root, and leaf extracts of C. latifolia were able to stimulate glucose uptake activity in 3T3-L1 adipocytes and L6 myotube cell lines [10].Additionally, the extracts were shown to have an insulinmimicking action that could activate insulin signaling cascade and enhance basal glucose uptake [10].C. latifolia shows potential in supporting health improvements that could be useful in the treatment of various diseases, such as diabetes, infammation, and cancer.Furthermore, the plant could serve as a valuable source of dietary supplementation that can contribute to overall health and vitality.
In this study, we aim to investigate the phytochemical contents and antioxidant and antidiabetic activities by chemical assays of C. latifolia leaves and C. latifolia roots.Additionally, the study also investigated the wound-healing activity of young and matured C. latifolia leaves on animal models at high and low doses.

Materials and Methods
In this paper, we evaluated the phytochemical contents and antioxidant activities of young (YCL) and matured (MCL) leaves of C. latifolia extracted in diferent solvents (n-hexane, dichloromethane, ethyl acetate, 80% (v/v) ethanol, 80% (v/v) methanol, and water), as well as ethanolic C. latifolia mixedaged leaf (CL) and C. latifolia root (CR) extracts.Subsequently, ethanolic CL extract was further assessed for its antidiabetic activity, while methanolic YCL and MCL extracts were investigated for their wound-healing activity on male Wistar rats.Te experimental design of this study is further illustrated in Figure 1.

Plant Collection and Sample Preparations. C. latifolia
leaves (matured and young) and roots were collected from Kampong Tenajor, Labi in Daerah Belait, Brunei Darussalam (4 °27′34.1″N,114 °28′09.2″E).Species were identifed and confrmed by UBD Botanical Research Centre (BRC) botanists, and a voucher specimen number B 038 880 was archived in Brunei Herbarium Sungai Liang. Figure 2 shows the C. latifolia group plantation found in Kuala Belait in Brunei Darussalam.
Te collected samples were washed with tap water to remove any remaining dirt and subsequently air-dried at room temperature for 4 weeks until a constant mass was reached.For the C. latifolia leaves, they were arranged according to their respective sizes, where large leaves between 60 and 120 cm were categorised as matured leaves, and smaller leaves in the range of 23-59 cm were labelled as young leaves (see Figure 3).Subsequently, the dried samples were pulverised using a kitchen blender (Zojirushi, BM-RSQ08) and sieved using 365-micron siever.Powdered samples were stored in desiccators at room temperature until further use.

Preparation of Crude Extracts by Soxhlet and Sequential
Extractions.Sequential extraction using n-hexane, dichloromethane, ethyl acetate, 80% (v/v) ethanol, 80% (v/v) methanol, and water was conducted on individual C. latifolia young and matured leaves.However, in the next part of the study, C. latifolia young and matured leaves were combined due to their limited amount and to simplify the collection process.

Sequential Extraction.
Diferent solvents of increasing polarity were used in sequential extraction.Te powdered samples of young and matured leaves of C. latifolia (50 g) were initially soaked in n-hexane (1 L) and shaken for 3 hours (SF1 Flask Shaker, Stuart Equipment, Stafordshire, United Kingdom), followed by 3 hours of sonication (Elmasonic E, E 180 H, Germany) at a constant temperature of 35 °C, and subsequently, fltered via vacuum fltration.With the residue obtained, the same procedure was repeated, following a predefned solvent sequence in the order of dichloromethane, ethyl acetate, 80% (v/v) ethanol, 80% (v/v) methanol, and water.Te use of diferent solvents was aimed to extract compounds with a range of polarities, from low to high.All extractions were conducted under a constant temperature of 35 °C, except for the solvent dichloromethane, which was maintained at 25 °C due to its lower boiling point.Consequently, from each fltrate, the solvent was evaporated using rotary evaporator and dried in an oven at 35 °C until a constant mass was obtained.Te dried crude extracts were stored in an airtight container and kept in desiccators at room temperature until further use.(1) Test for Alkaloids.Te presence of alkaloids was tested according to the method described by Tadesse et al. [11] with minor modifcations.Each extract (20 mg) was dissolved in 1% (v/v) hydrochloric acid (8 mL) and fltered.Te fltrate attained was then mixed with 1 mL of potassium bismuth iodide solution (Dragendorf's reagent), where formation of orange red precipitate shows the presence of alkaloids.
(2) Test for Saponins.Te extracts were screened for the presence of saponins using the methods outlined by Auwal et al. [12].Each extract (50 mg) was mixed with 1 mL of distilled water in a test tube and the mixture was shaken vigorously.In the presence of saponins, formation of stable frothing would be observed.
(3) Test for Tannins.Te screen test for tannins was investigated with the procedures detailed by Auwal et al. [12].Each extract (50 mg) was dissolved in distilled water (1 mL) before drops of 5% (w/v) ferric chloride were added to the mixture.Te formation of black or blue-green precipitate shows that tannins are present.
(4) Test for Terpenoids.Te extracts were each tested for the presence of terpenoids according to the method by Tadesse et al. [11].Te test, namely, Salkowski test, was conducted by mixing the extract (50 mg) together with 1 mL of chloroform, followed by 1 mL of concentrated sulfuric acid, which was poured slowly along the sides of the test tube.
(5) Test for Glycosides.For identifcation of glycoside, the Keller-Killiani test was performed using the methods described by Gul et al. [13] with slight modifcations.Each extract (50 g) was mixed with 5 mL of distilled water, followed by an addition of 2 mL of glacial acetic acid, together with 2-3 drops of ferric chloride and 1 mL of concentrated sulfuric acid along the sides of the test tube.A positive result for the presence of glycoside would give a formation of brown or velvet ring.
Te mixture was incubated for 30 minutes and subsequently measured for its absorbance at wavelength 725 nm using a single beam UV-visible spectrometer (GENESYS ™ 20S, Termo Fisher Scientifc Inc., USA).Te standard calibration curve for gallic acid (0-100 mg/L) was prepared in the same manner and the TPC of extracts was attained using equation (1), where the results were expressed as gallic acid equivalence (GAE) in mg/g of extract.
TPC in mg GAE g of extract � [Gallic acid] in mg/L × volume of extract(L) mass of extract (g) . (1)

Total Flavonoid Content.
Total favonoid content (TFC) was determined using the aluminium colorimetric method described by Salim et al. [15] with some modifcations.Te extract solution (0.5 mL) was mixed with 10% (w/v) aluminium chloride (0.1 mL), 1 M potassium acetate (0.1 mL), absolute ethanol (1.5 mL), and distilled water (2.8 mL).Te mixture was incubated for 30 minutes, and the absorbance was measured at wavelength 415 nm using a single beam UV-visible spectrometer (GENESYS ™ 20S, Termo Fisher Scientifc Inc., USA).Subsequently, the TFC of extracts was obtained from the calibration curve of quercetin (1-100 mg/L), using equation (2), and the results were expressed as quercetin equivalence (QE) in mg/g of extract.
TFC  [16] with some modifcations.Briefy, a stock solution of 50 mg/L DPPH was made fresh with absolute methanol prior to analysis.A standard gallic acid was prepared in concentrations between 0.5 mg/L to 10 mg/L, whereas the extract solutions were made into several concentrations between 10 and 100 mg/L.Subsequently, the DPPH stock solution (200 µL) was added into each of the standard solutions and extract samples (25 µL) and further kept in the dark for 30 minutes.Te absorbance was then taken using a microplate reader (BIOBASE, BK-EL10C, China) at 517 nm against methanol as the negative blank.Te percentage DPPH radical inhibition was calculated using the following equation.

Percentage inhibition of DPPH radical (%) �
Te IC 50 was obtained from the calibration curve of percentage inhibition against the concentrations of each extract or standard.Subsequently, the DPPH radical scavenging activity (RSA) was determined based on equation ( 4) and the results were expressed in mg gallic acid equivalent (GAE)/g extract.

DPPH
Te experiment was conducted in triplicate and the data obtained are expressed in mean ± standard deviation.
Te Scientifc World Journal 2.5.2.α-Amylase Enzyme Activity.Amylase activity of the individual extracts was determined using a colorimetric method based on a commercialised amylase assay kit (catalog no.MAK009, Sigma-Aldrich, Merck, USA).Sample extracts and positive control, acarbose (200 mg/L), were diluted in Amylase Assay Bufer from the kit, and nitrophenol standards of varying concentrations of 0 (blank), 4, 8, 12, 16, and 20 nmol/well standards were prepared to obtain a linear range of calibration curve.Prior to assay reaction, Master Reaction Mix was made ready by mixing an equal ratio of Amylase Assay Bufer (50 µL) and Amylase Substrate Mix (50 µL) and then added into individual wells of 96-well plate containing 50 µL of the extracts, standards, and positive control.Te initial absorbance at 405 nm was measured [(A 405 ) initial ] using a 96-well microplate reader (BIOBASE, BK-EL10C, China), and the absorbance was recorded every 5 minutes for 35 minutes.Te sample with the highest change in absorbance (calculated using equation ( 7)) was then used to determine the amylase activity (using equation ( 8)).
Amylase activity Te number of moles of nitrophenol in the sample was evaluated from the standard calibration curve of nitrophenol.Te amylase activity of the extracts and positive control was expressed as milliunit (mU), which is equivalent to nmol/min/mL.One unit of amylase refers to the amount of amylase that links ethylidene-pNP-G7 to produce 1.0 μmol of p-nitrophenol per minute.

Animal Studies.
Ethanolic extract of C. latifolia mixedage leaves was investigated for its antihyperglycemic efects on alloxan-induced diabetic rats.Additionally, methanolic extracts of young and matured C. latifolia leaves were evaluated for their wound-healing potential on male Wistar rats.

Antihyperglycemic Activity on Animal Model.
Antihyperglycemic study was conducted based on the protocol described by Hazirah Matusin et al. [19].In this preliminary study, C. latifolia leaf extract was chosen to evaluate its capacity to lower down blood glucose level (BGL) and changes in body weight percentage of alloxaninduced diabetic rats.
(1) Experimental Animals.Male Wistar rats weighing 160-200 g and aged between 8 and 10 weeks old from the animal house of Universiti Brunei Darussalam were used.Te animals were housed in polypropylene cages that were cleaned regularly and were provided ad libitum access to water and standard pellet rat diet (Altronim, Germany).Te procedures in this antihyperglycemic experimental study were approved by the University Research Ethics Committee (Ref File: UBD/FOS/E2.(j)).Upon completion of experimental procedures, animals were euthanised using carbon dioxide asphyxiation.Te carbon dioxide was gradually flled into the euthanasia chamber at a fow rate between 30 and 70% of the chamber volume per minute or 400-1000 ppm, depending on the size of the rats.Animals were kept in the chamber for a few minutes until death was confrmed by checking their respiratory arrest or observing their fxed and dilated pupils.
(2) Administration of Alloxan.Animals were abstained from food for 16 hours and were only provided with water before administration of alloxan.0.9% (w/v) saline was used to dissolve the alloxan monohydrate (Sigma-Aldrich, United Kingdom) and a single dose of 120 mg/kg body weight (b.w.) was administered through the intraperitoneal cavity part of the animal to instigate diabetes.Teir blood glucose levels were observed 72 hours after administration of alloxan and animals with readings above 15.0 mmol/L were deemed to be diabetic and were used in the antihyperglycemic study.
(3) Experimental Groups of Antihyperglycemic Study.Tis research project is composed of three main groups, where each group consists of six animals: normal untreated control (n � 6), diabetic control group (n � 6), and treated with C. latifolia leaf extract group (n � 6).Normal untreated control consists of healthy normal male Wistar rats, while diabetic control involves animals induced with 120 mg/kg b.w. of alloxan and not treated with sample extract.Animals that showed high readings of blood glucose 72 hours after alloxan administration were chosen for this study and only received normal diet throughout the experiment.For the treatment group, oral gavage administration of C. latifolia leaf extract at 250 mg/kg b.w. was carried out on the animals twice a week over a period of 14 weeks.Te dosage of extract was determined based on the method by Hazirah Matusin et al. [19].Subsequently, body weight and blood glucose reading of each rat were recorded on a weekly basis.An acute toxicology observation on alloxan-induced animals treated with 250 mg/kg b.w.C. latifolia leaf extract demonstrated no signs of morbidity or mortality throughout the experiment, which suggested the dosage was safe to use.

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Te Scientifc World Journal (4) Blood Glucose Level and Body Weight Percentage.Te animals in this study were regularly measured for their blood glucose level in mmol/L using blood glucose test strips and a glucometer (FreeStyle Optium, Abbot Diabetes Care Ltd.).
Measurement of blood glucose level was obtained by drawing blood from the tail tips using a single touch strip.Nonfasting animals of normal control, diabetic control, and treatment groups were regularly recorded for their blood glucose levels for 14 weeks, for which readings above 11.1 mmol/L were considered diabetic.Simultaneously, body weights of the animals were also taken once a week using a digital weighing machine (BX22KH Model, Shimadzu Corporation) and the data obtained were expressed in body weight percentage (%).
(5) Intraperitoneal Glucose Tolerance Testing (IPGTT).An IPGTT test was conducted on all the animals in normal control, diabetic control, and treatment groups after the fnal experimental period had ended.Te animals were required to fast for 16-18 hours and subsequently were measured for their blood glucose reading; this time point was defned as 0 min.Each animal in every group was injected with a single dose of 2 g/kg b.w.D-glucose and their blood glucose levels were monitored at a time interval of 15, 30, 60, 90, and 120 minutes.Blood glucose readings were expressed as mmol/L.

Wound-Healing Activity on Animal Model.
Wound-healing activity was conducted based on the methods described by Shafe et al. [20].Methanolic young and matured leaves of C. latifolia leaves were investigated for their wound-healing efects on male Wistar rats.Tis preliminary study measured the percent wound contraction of the animals after topical treatment of the extracts on alternate days for 14 days.
(1) Experimental Animals.In this research project, adult male Wistar rats of 8-10 weeks old, weighing about 200-400 g, from the animal house of Universiti Brunei Darussalam were used.Te rats were individually placed in clean polypropylene cages and provided ad libitum access to food and water.Experimental procedures were approved by the University Research Ethics Committee (Ref fle: UBD/ FOS/N(1)).After the experiments were completed, animals were euthanised using carbon dioxide asphyxiation.Te carbon dioxide fow rate was between 30 and 70% of the euthanasia chamber volume per minute or between 400 and 1000 ppm, depending on the size of the rats.Animals were maintained in the chamber for several minutes until death was confrmed by observing their fxed and dilated pupils and respiratory arrest.
(2) Wound Excision.Prior to excision procedure, the rats were individually anaesthetised with diethyl ether by inhalation anaesthesia.A few drops of diethyl ether were applied onto a cotton wool and then placed at the bottom of a transparent acrylic jar.Subsequently, the animals were exposed to the diethyl ether without direct contact between the cotton wool and the animal.Te level of anaesthesia by exposure of diethyl ether was assessed by observing the animals' ability to make voluntary movements, such as extending their legs in response to stimuli.After ensuring deep anaesthesia of the animals, wound excision was performed.Te dorsal thoracic region was shaved using an electric shaver (Surker, HD 8202, UK) and then cleaned with absolute ethanol before excising the skin (10 mm × 10 mm) using forceps and pointed scissors (see Figure 4).Te wound area was further treated with saline, and the rats were then moved to a well-ventilated area to recover from the anaesthesia.Topical application of prepared ointments on wound area was done once in every 2 days intervals for a period of 14 days for each treatment group.
Furthermore, extraction yield of C. latifolia mixed-age leaves and C. latifolia roots extracted using Soxhlet extraction in 80% (v/v) ethanol is shown in Figure 5(b).Our data showed CL extract (14.75 ± 0.20 g/100 g) produced a slightly higher yield compared to CR extract (11.44 ± 0.38 g/100 g), suggesting an insignifcant diference between the yields of two extracts.Additionally, Soxhlet extraction demonstrated a better extraction of ethanolic leaves compared to sequential method, with the ethanolic Soxhlet yield (14.75 ± 0.20 g/100 g) being roughly two times more than that of the sequential ethanolic leaf extracts (MCL, 7.25 ± 0.14 g/100 g, and YCL, 8.47 ± 0.20 g/100 g).Te qualitative analyses to screen the presence of phenolic and favonoid were not conducted in this study.However, a quantitative study of total phenolic and favonoid contents was performed, where the results are shown in Section 3.2.2.

Quantitative Total Phenolic and Flavonoid Contents.
Te quantifcation of TPC and TFC in the sequential extracts of YCL and MCL and ethanolic extracts of CL and CR is presented in Figure 6.
A comparison study of TPC and TFC between sequential extracts at 500 mg/L showed high phenolic and favonoid contents in EtOH-YCL (4.02 ± 0.004 mg GAE/g of extract and 11.083 ± 0.004 mg QE/g extract, respectively).Tis was followed by EtOH-MCL with TPC of 3.087 ± 0.002 mg GAE/ g of extract and TFC of 7.387 ± 0.004 mg QE/g extract (see Figures 6(a) and 6(c)).Te results are supported by a statistical analysis which showed a signifcant diference of phenolic and favonoid contents (p < 0.05) between YCL and MCL.
While young leaves overall showed a greater TPC and TFC, the next part of the study chose to combine the young and matured leaves of C. latifolia for the purpose of simplifying the plant collection process.In the TPC analysis for CL and CR extracts at 1000 mg/L, CR revealed a higher value of 359.80 ± 0.01 mg GAE/g of extract compared to CL (141.19 ± 0.01 mg GAE/g of extract).
However, our TFC analysis demonstrated a contrasting result where CR exhibited a lower reading of 17.89 ± 0.00 mg QE/g extract, while CL yielded 68.70 ± 0.00 mg QE/g extract (Figure 6(d)).

Antioxidant Activities.
Te antioxidant activity was evaluated by DPPH radical scavenging activity and ferric reducing antioxidant power assays.Figure 7 shows the results for the DPPH RSA and FRAP of sequential extracts of young and matured C. latifolia leaves and ethanolic extracts of C. latifolia leaves and C. latifolia roots.
Among the examined sequential extracts, ethanolic extracts showed the highest DPPH radical scavenging activity with YCL having RSA of 143.4 ± 2.1 mg GAE/g of extract and MCL at 127.2 ± 10.8 mg GAE/g of extract (see Figure 7(a)).Te antioxidant activity by FRAP assay also  Te Scientifc World Journal found ethanolic extracts to be the most efective in reducing Fe 3+ /ferricyanide complex to ferrous Fe 2+ by donating free electrons (YCL, 2.120 ± 0.067 mg•TE/g extract, and MCL, 1.868 ± 0.011 mg•TE/g extract) (see Figure 7(c)).Simultaneously, ethyl acetate, methanolic, and aqueous extracts demonstrated promising antioxidant activity in DPPH radical scavenging activity and FRAP assays, suggesting the efcacy of high polar solvents in extracting antioxidants.
Our fndings on the antioxidant activities of sequential extracts of YCL and MCL show consistency with our TPC and TFC results which could suggest that phenolic and favonoid compounds are the primary antioxidants.Tis was supported by a statistical analysis which resulted in a positive correlation of DPPH RSA and FRAP with TPC and TFC (see Table 2).
In the next study, we evaluated the antioxidant activity of CL and CR extracts.Results demonstrated that CR exhibited the most promising antioxidant activity with the highest  Te Scientifc World Journal RSA of 34.06 ± 0.14 mg•GAE/g extract and greatest FRAP activity at 512.43 ± 0.05 mg•TE/g extract.Tis was followed by CL extract with RSA of 7.99 ± 0.09 mg•GAE/g extract and FRAP of 177.29 ± 0.33 mg•TE/g extract.Our fndings on the antioxidant activities of ethanolic CL and CR extracts showed moderate correlation with TPC (r � 0.76), but a negative correlation with TFC (r � −0.16) (see Table 2).CR extract with a lower value of TFC (17.89 ± 0.00 mg•QE/g extract) presented a greater antioxidant activity, while CL with the higher TFC (68.70 ± 0.00 mg•QE/g extract) showed a lower RSA and reducing power as shown in Figures 7(b  Of all the individual extracts, CR exhibited the highest ∝ -glucosidase inhibitory activity at 74.68 ± 2.14%, which is comparable with that of a marketed antidiabetic drug, acarbose (96.21 ± 2.03%) (see Figure 8(a)).Meanwhile, CL extract showed a lower ∝ -glucosidase inhibition activity with a percentage inhibition of 32.40 ± 1.13%.
Furthermore, the data for ∝ -amylase enzyme activity are expressed as nmol/min/mL (milliunit (mU)), where one unit of amylase refers to the amount of amylase that links ethylidene-pNP-G7 to produce 1.0 μmol of p-nitrophenol Te Scientifc World Journal 12 Te Scientifc World Journal

Antihyperglycemic Activity on Animal Model.
In this present study, ethanolic C. latifolia mixed-age leaf extract was investigated for its efect on the blood glucose levels of alloxan-induced male Wistar rats, followed by the evaluation of body weight percentage and intraperitoneal glucose tolerance test (IPGTT).
(1) Blood Glucose Level and Body Weight Percentage.Blood glucose readings of animals in normal control, diabetic control, and CL-treated groups are displayed in Figure 9. Te normal untreated group showed consistent blood glucose levels between 5.05 ± 0.80 to 6.02 ± 0.59 mmol/L, while alloxan-induced diabetic control group continued to show high blood glucose concentrations between 20.76 ± 1.46 and 23.52 ± 0.99 mmol/L.Te results obtained from these control groups are used as reference for normal and diabetic blood glucose levels to compare with our fndings on CLtreated group.As a result, BGL of the extract-treated diabetic rats demonstrated a signifcant decrease (p < 0.05) at the frst 5 weeks after induction of alloxan and treatment with 250 mg/kg b.w.CL extract, attaining a reading of 7.6 ± 1.21 mmol/L by the fnal week of treatment.Subsequently, the body weights of the animals were monitored on a weekly basis until the end of the experimental period.Te data are represented as percentage change in body weight as displayed in Table 3. Diabetic animals receiving treatment of 250 mg/kg b.w.C. latifolia leaf extract were shown to have a signifcant body weight gain (p < 0.05), reaching a percentage change of 45.83 ± 7.33% at week 14.Te results were comparable to that of the normal control group which showed a body weight change of 32.50 ± 3.40% by the end of the experimental period.Meanwhile, the alloxan-induced diabetic group displayed a reduction in body weight of −10.16 ± 5.35%, which is a characteristic sign of diabetes. (

2) Intraperitoneal Glucose Tolerance Testing (IPGTT).
IPGTT was also carried out on the animals of normal control, diabetic control, and CL-treated groups at the end of their 14-week experimental period, where their fasting blood glucose levels were measured to evaluate their glucose tolerance.Te results are displayed in Figure 10, where changes in blood glucose levels are monitored at a time interval of 15, 30, 60, 90, and 120 minutes.Te initial mean BGLs at 0 minutes for fasting animals in normal control and CL treatment groups were 4.12 ± 0.48 and 4.43 ± 0.39 mmol/L, respectively, which are within the range of normal fasting blood glucose concentrations (3.9 to 5.6 mmol/L).Meanwhile, the initial fasting blood glucose level of diabetic control group was 21.12 ± 1.59 mmol/L which exceeded the diabetic threshold value of 7.0 mmol/L.IPGTT of CL treatment group was comparable with that of normal control group, where the fnal BGLs at 120 minutes were 4.88 ± 0.90 mmol/L and 4.82 ± 0.58 mmol/L, respectively.Moreover, the results were signifcantly lower than the blood glucose readings of diabetic rats (27.4 ± 0.52 mmol/L at 120 minutes).

Wound-Healing Activity on Animal Model.
In our preliminary wound-healing study, methanolic extracts of young (Me-YCL) and matured C. latifolia (Me-MCL) leaves were investigated for their wound-healing activity on male Wistar rats of 8-10 weeks old.Te wound area of treated animals was topically applied with high dose 50% (w/w) Me-YCL, high dose 50% (w/w) Me-MCL, low dose 10% (w/w) Me-YCL, and low dose 10% (w/w) Me-MCL.Subsequently, wound contractions of the animals were measured on Day 3, 5, 7, 10, 12, and 14 (see Figure 11).High dose 50% (w/w) Me-YCL showed a prominent activity in wound healing when compared to control group 1 (untreated) at the beginning of the study.When comparing the results between untreated Te Scientifc World Journal control group and the 50% (w/w) Me-YCL, a diference in wound contraction on Day 3, 5, and 7 was evident, where untreated group reported the measurements of 3.8 ± 0.7%, 16.5 ± 0.5%, and 52.0 ± 0.6%, respectively, whereas 50% (w/w) Me-YCL reported the measurements of 9.1 ± 1.3%, 30.7 ± 0.5%, and 67.8 ± 0.0%, respectively.Additionally, 50% (w/w) Me-MCL also showed promising wound-healing progression on Day 3, with a wound contraction of 8.6 ± 2.4%, while that of untreated control group was at 3.8 ± 0.7%.At the end of the experimental study (between Day 12 and Day 14), all the experimental groups had eventually reached more than 90% of wound contraction.However, none of the treated groups showed a prominent diference in wound contractions when compared with the untreated control group.
Table 4 shows the physical observation of the woundhealing progression for all the groups on Day 1, 3, 5, 7, 10, 12, and 14.Scab formation began to occur on Day 3 for all the experimental groups (including control).Meanwhile, detachment of these scabs can be seen on Day 10 for 50% (w/ w) Me-MCL, 50% (w/w) Me-YCL, 10% (w/w) Me-MCL, and 10% (w/w) Me-YCL, while the scabs in the control groups were retained until Day 12. Additionally, wound size of animal treated with 50% (w/w) Me-YCL appeared to be the most diminished on Day 10 compared to the rest of the experimental groups, which shows consistency with its reported wound contraction seen in Figure 11.14

CL-Treatment group
Te Scientifc World Journal compared to low polar solvents, with 80% (v/v) methanol being the best, followed by 80% (v/v) ethanol and water.Our results show consistency with a previous study by Nur et al. [21] which reported high polar solvent, specifcally 70% (v/v) ethanol, to be efective in dissolving polar compounds and producing high extraction yield of C. latifolia leaf extract.In another fnding by Ooi et al. [22], extraction yield of C. latifolia rhizome aqueous extract was the highest compared to low polar solvents such as hexane and ethyl acetate.Solvents with high polarity such as ethanol, methanol, and water are commonly used in the extraction of natural products to extract polar compounds and often result in high percentage yield [23].Additionally, hydro-alcohols are also common extraction solvents which are efcient in extracting a wide range of compounds including hydrophilic (watersoluble) and lipophilic (fat-soluble) substances [24].Te polarity of ethanol solvent is more consistent in dissolving polar components such as phenolic compounds, glycosides, steroids, alkaloids, polysaccharides, and phospholipids [21,24].Meanwhile, water as a solvent system is efective in extracting water-soluble components such as saponins, tannins, terpenoids, and anthocyanins [25].
Diferent extraction methods can also afect the percentage yield, as our results demonstrated Soxhlet method to give a higher yield of ethanolic extracts compared to sequential extraction.Te use of high temperature in the Soxhlet method allows a reduction in water density and viscosity, further increasing the mass transfer of the solvent into the sample matrix [26].However, increasing the temperature to a certain point can also cause the solution to saturate, preventing the dissolution of solids, and subsequently, reducing the extraction yield [26]. Tis was evident in the study by Zabidi et al. [26], where the optimum temperature for the extraction of C. latifolia roots was at 180 °C and as the temperature increased to 200 °C, its percentage yield decreased.Based on our study, extraction of C. latifolia plant using the Soxhlet method and high polar solvents such as 80% (v/v) ethanol, 80% (v/v) methanol, and water result in the best extraction yield.

Phytochemical Contents by Chemical Assays.
In the present study, phytochemicals such as tannins, saponins, glycosides, and terpenoids were found in all the extracts (refer to Table 1).Tese secondary metabolites are known for their protective role against diseases and infections as diabetes, cancer, and cardiovascular diseases.Previous studies have found C. latifolia leaves to contain several phytochemicals such as cinnamic acid, berberine, and glycosides which may have played a part in the scavenging of free radicals and antidiabetic activity [27].
Furthermore, our fndings showed that using diferent extraction solvents resulted in diferent phytochemical profles, with diverse compositions and concentrations of active compounds.Generally, nonpolar solvents like nhexane and dichloromethane have the tendency to extract nonpolar compounds, whereas polar solvents such as methanol and water would most likely extract polar substances.Tis accounts for the observed diference in phytochemical response, and as expected, the C. latifolia leaves Te Scientifc World Journal  Aside from extraction solvents, the maturity of leaves was also found to have an impact on the extraction of phytochemicals.In this study, young leaves of C. latifolia were shown to contain a higher content of phenolic and favonoid compounds compared to matured leaves.Tis is supported by a previous fnding which reported the decrease of favanols and phenolic acid content upon leaf aging [28].Nevertheless, it is also demonstrated that some valuable compounds such as rutin were found accumulated in matured leaves which could benefcial for treating infammation, lowering cholesterol levels, and improving blood circulation [28].Terefore, both young and matured can be valuable components of a diverse and nutritious diet which can help in maintaining the overall health and wellbeing of humans.
In the next study, mixed-age C. latifolia leaves and C. latifolia roots in 80% (v/v) ethanol were analysed.Our results showed that CR had the most promising TPC compared to CL extract.Tere has been a previous study that revealed CR extract to contain phenolic compounds including scandenin, pomiferin, phloridzin, and mundulone [26].Te presence of these phenolics was described to play a key role in its antioxidant [29] and antidiabetic [30] activities.However, there are still limited studies that have reported on the phenolic content of C. latifolia root extract and further extensive investigation is needed to assess the extent of the biological capacities of these chemical compounds.
Te discrepancy in TPC results between CR and CL could be due to various reasons, including environmental biotic and abiotic stresses [29].Additionally, the use of heat in the Soxhlet extraction process may have also resulted in the loss of some important volatile phenolic compounds in CL extract.Terefore, modifcations to the study such as employing diferent extraction methods and varying the ratio for water-ethanolic solvents could be worthwhile to extract other potential bioactive phytochemicals that could exert various biological activities such as antidiabetic, antiinfammatory, wound-healing, and antimicrobial activities.
Interestingly, our fndings revealed TFC of CR to be lower than CL (see Figure 6(d)), indicating that the majority of phytochemical compounds present in CR extract may be composed of phenolics rather than favonoid compounds.However, favonoid compounds existing in CR could still impart useful medicinal properties.For instance, Ullah et al. [31] described that favonoids can serve as exogenous antioxidants, which can form free radicals to less reactive species through mechanisms such as suppressing nitric oxide synthase activity, inhibition of xanthine oxidase activity, modulation of channel pathways, or interacting with other enzyme systems.Quercetin, a potent antioxidant favonoid, has been widely reported for its versatile protective abilities, including anti-infammatory, antihypertensive, and antiobesity activities [32].Additionally, the active compound has shown neuroprotective efects through its ability in suppressing neuroinfammation and promoting memory, learning, and cognitive functions [33].As for CL extract, its greater TFC is consistent with a previous study by Umar et al. [29], of which it was indicated that favonoids are generally accumulated in the aerial parts of the plant to provide protection against pathogenic attacks or solar radiation.
Te phytochemicals found in CL and CR extracts possess signifcant potential in supplementing dietary needs that could contribute to the enhancement of overall health and in the treatment of various chronic diseases or health conditions [34].Further extensive studies on these bioactive compounds are needed to gain more understanding into their properties, potential application, and mechanism of actions.Tis includes investigating the bioactivity of their isolated active compounds and elucidating the molecular pathways through which these compounds impact the cellular processes.

Antioxidant Activities.
Antioxidants play a crucial role in the protection of cells against free radicals that are implicated in the pathogenesis of many diseases such as cancer, infammation, DNA damage, lipid peroxidation, cardiovascular disease, and diabetes [35].Many phytochemicals have antioxidant properties that can quench free radicals, chelate catalytic metals, and subsequently terminate chain reactions and inhibit oxidation reactions [35,36].Consequently, these antioxidant agents can stimulate the immune system, block the formation of carcinogen, and prevent the production of estrogen implicated in breast cancer [36].
Among the sequential extracts, ethanolic YCL and MCL extracts exhibited the most efective DPPH radical scavenging activity and ferric reducing power, while hexane and dichloromethane extracts were the lowest.Our fndings showed consistency with our TPC and TFC results which could suggest that the polar polyphenol compounds are the primary antioxidants that are responsible in scavenging free radicals and reducing ferric ions.Another observation from Figures 7(a) and 7(c) can be seen where YCL demonstrated a greater antioxidant activity compared to MCL.Tis is supported by the statistical analysis that showed a signifcant diference of antioxidant activity (p < 0.05) between the young and matured leaves.Furthermore, our results agree with a previous fnding, which reported that immature leaves overall exhibited higher antioxidant capacities through DPPH, FRAP, 2,2′-azino-bis-(3-ethylbenzothiazoline-6-sulfonic acid) diammonium salt (ABTS), and nitric oxide chemical assays [37].In addition, the study by Ti and Hwang [38] described that during leaf maturation, diferent oxidative metabolism and antioxidative strategies in plant tissues take place.Teir fndings reported younger Aronia leaves to have higher antioxidant activity which is consistent with our results on YCL [38].
While ethanolic YCL and MCL extracts showed positive results, our next study demonstrated ethanolic CR extract to exhibit a more prominent radical scavenging activity and ferric reducing antioxidant power.Despite CR's very low TFC, it can be hypothesised that its phenolic compounds were the major contributors in scavenging DPPH radicals Te Scientifc World Journal and reducing ferric ions.In a previous study by Zabidi et al. [26], the presence of phenolic compound, hydroquinone, in C. latifolia root extract was suggested to have contributed to its strong antioxidant activity.Te active compound and its derivatives contain hydroxyl groups that can exert a strong radical scavenging activity between 93% and 97%, comparable to a known antioxidant, ∝ -tocopherol (95%) [39].Moreover, cinnamic acid in the extract may have also played a major role in the antioxidant activity, where its hydroxyl groups can donate hydrogen or electron, further neutralising free radicals [40].
Overall, ethanolic extracts of C. latifolia leaves and C. latifolia roots potent antioxidant capacity that is important in preventing degenerative diseases such as cancer, diabetes, infammation, and obesity.Tis activity may be related to the presence of active phenolic compounds in the extracts that could absorb and stabilise highly reactive free radicals.However, more studies are still needed to further validate this study.Future recommendations which can potentially result in the extraction of more potent antioxidants may include the use of other polar solvents, extraction methods, and diferent proportions with aqueous solvent.

Antidiabetic Activity by Chemical Assays
. Te antidiabetic potential of C. latifolia leaves and C. latifolia root extracts was investigated via ∝ -glucosidase and ∝ -amylase inhibitory assays.Te extracts were evaluated on their capacity to inhibit the activity of the two enzymes that are involved in the digestion of carbohydrates and starch which usually causes the increase of postprandial (after-meal) hyperglycemia in diabetic patients [41].Postprandial hyperglycemia is characterised by an elevated blood sugar concentration following a meal, and in T2DM, their impaired insulin response leads to a delayed uptake of glucose by the cells, making it difcult to maintain near normal blood glucose readings [42].Terefore, inhibiting ∝ -glucosidase and α-amylase is a strategy to control blood sugar levels by regulating the digestion and absorption of carbohydrates in the digestive system.
Te present study demonstrated a promising antidiabetic activity of CL and CR extracts through ∝ -glucosidase inhibition and ∝ -amylase enzyme activities.Among the individual extracts, CR extract stood out the most by showing marked ∝ -glucosidase and ∝ -amylase inhibition activities that were similar to that of a marketed antidiabetic drug, acarbose.Tis indicated that CR have promising active components that can efectively inhibit these two enzymes which could further help in regulating postprandial hyperglycemia and lowering the risk of developing diabetes [43].Te extract's phenolic and favonoid contents and phytochemical compounds such as saponins, tannins, terpenoids, and glycosides may have individually or synergistically been involved in the mechanism of action [44].Tis agrees with the study by Umar et al. [29] which demonstrated a signifcant correlation between high phenolic content and ∝ -glucosidase inhibition activity.Te functional group, confguration, substitution, and number of hydroxyl groups of these phytochemical constituents are the key elements in the attachment to the binding sites of ∝ -glucosidase and ∝ -amylase enzymes [45].Tis leads to a more regulated rate at which carbohydrates and starch are broken down and subsequently would result in the improvement of insulin sensitivity and glucose uptake [46,47].
On the other hand, CL showed less prominent activity in both ∝ -glucosidase inhibition and ∝ -amylase enzyme assays than CR.Nonetheless, CL's ability to hinder the action of ∝ -glucosidase and ∝ -amylase enzymes could still have potential in suppressing the breakdown of carbohydrates in the small intestine and delaying the glucose uptake [41].Its considerable antidiabetic properties could be greatly benefcial in the development of medication for the treatment of diabetes.Moreover, the dosage of extract may play a key factor in inducing much greater ∝ -glucosidase inhibition and ∝ -amylase enzyme activities.Increasing the concentration of CL extract may possibly lead to an enhanced outcome; therefore, more studies are needed to determine the right dosage.

Animal Studies.
In this present study, we evaluated the antihyperglycemic activity of ethanolic C. latifolia leaf (mixed-age) extract on alloxan-induced male Wistar rats.C. latifolia mixed-age leaves were selected due to its abundant extraction yield which was sufcient to achieve data for this experimental study.Additionally, there are limited studies conducted on the antihyperglycemic efects of ethanolic C. latifolia leaf extract; therefore, fndings from our study could be considered novel.Based on our fndings, C. latifolia leaf extract at 250 mg/kg b.w. was able to reduce the blood glucose levels of diabetic rats to normal within 14 weeks.BGL of the extract-treated diabetic rats demonstrated a signifcant decrease (p < 0.05) at the frst 5 weeks after induction of alloxan and attained a reading of 7.6 ± 1.21 mmol/L by the fnal week of treatment.Te results obtained were comparable with those of normal control group by the end of the experiment (see Figure 9), which implies that the treatment of 250 mg/kg b.w.C. latifolia leaf extract was efective in exerting glucose-lowering efects, indicating its antihyperglycemic potential.
Subsequently, the animal body weights of CL treatment group had signifcant improvement compared to the animals in alloxan-induced diabetic control.Te percentage body weight gain of the treatment group was highly comparable to that of normal control by week 14.Terefore, the extract showed efcacy towards ameliorating the body weight gain of diabetic animals, suggesting its ability to stimulate insulin secretion by inhibiting lipolysis, proteolysis, and glycogenolysis [48].Moreover, C. latifolia leaf extract showed promising protective efects against diabetes-inficted weight loss, which is consistent with the signifcant reduction of blood glucose levels.Tese efects may be attributed to the presence of phytochemicals such as phenolic acids, favonoids, terpenoids, tannins, and saponins which have been proven to exhibit antihyperglycemic properties.Tus, our phytochemical screening (see Table 1) has shown C. latifolia leaf extract to possess these active components which may 18 Te Scientifc World Journal facilitate in the synergistic interaction on molecular targets, leading to measurable therapeutic efects.Additionally, their ameliorative efects on blood glucose regulation indicated that the extract was efective in regenerating β-cells, stimulating insulin secretion, and maintaining normal glucose homeostasis [49,50].Simultaneously, the presence of antioxidants in the extract may help in reducing oxidative stress, further preventing major complications of diabetes such as cardiovascular disease, kidney disease, and nerve damage [51].Furthermore, intraperitoneal glucose tolerance testing (IPGTT) was carried out on all the experimental groups after their 14 weeks of treatment, of which their fasting blood glucose levels were measured to evaluate their glucose tolerance.the study, CL-treated diabetic rats showed marked improvement as their blood glucose levels reached 4.88 ± 0.90 mmol/L 120 minutes after being administered with 2 g/kg b.w.D-glucose.Te results obtained were comparable with those of normal untreated control throughout the 120 minutes IPGTT experiment, indicating that CL-treated diabetic animals may have achieved normal bodily functions after the 14 weeks of treatment.Meanwhile, BGL readings of diabetic animals maintained above the normal threshold, indicating its impairment in insulin secretion and regulating blood sugar levels.
Tere have been previous fndings that reported on the antidiabetic properties of C. latifolia plant.A study by Ishak et al. [30] revealed aqueous extract of C. latifolia fruit: root at 200 mg/kg b.w.signifcantly decreased the glucose level of streptozotocin-(STZ-) induced diabetic rats.Te results found that the extract was efective in preventing further of β-cells and improved insulin and adiponectin secretions that contributed to the decrease of glucose and lipid levels of the animals [30].A previous study by Ooi et al. [52] has described that the bioactive compound, curculigoside, may possess antidiabetic properties from its high efcacy in glucose transport activity via stimulation of glucose transporter 4 (GLUT4) levels at the plasma membrane in 3T3-L1 adipocytes.Additionally, cinnamic acid in C. latifolia was also indicated to contribute to facilitating insulin secretion by activating the voltage-dependent Ca 2+ channels, inducing an increase in Ca 2+ and closure of ATPsensitive K + channels, without causing membrane depolarisation [27,53].Furthermore, C. latifolia has been revealed to contain sweet proteins known as curculin and neoculin [54].Tese proteins were found to be 500 to 9000 times sweeter than sucrose by weight, and thus they have been used as low-calorie sweeteners and taste modifers [55].Teir nature as sugar substitutes may have potential as antidiabetic agents.However, no studies have been conducted to confrm the notion; therefore, further studies would be worthwhile to fnd out their antidiabetic capacity.
Our fndings could suggest C. latifolia leaves as a potential source for the development of food supplements, which could improve insulin sensitivity, glucose metabolism, and overall health and well-being.However, there is still a lack of understanding on the mechanism of action and the role of phytochemical constituents underlying the bioactivity of the leaf extract as there are very limited studies in this area.Extensive studies involving microscopic anatomy and immunohistochemical analysis on the body organs of animals afected by diabetes such as pancreas, liver, and kidney could therefore ofer more information and better understanding of its pathology.
Diabetic patients typically have a higher risk of experiencing complications related to wound healing compared to nondiabetic individuals.Consequently, the next study investigated the wound-healing efects of methanolic young and matured C. latifolia leaf extracts on male Wistar rats.Based on our results, high dose 50% (w/w) Me-YCL resulted in an enhanced wound repair process due to its higher content of phenolic and favonoid compounds.Te wound contractions between untreated control group and the 50% (w/w) Me-YCL were signifcant (p < 0.01) on Day 3, 5, and 7, where untreated group resulted in 3.8 ± 0.7%, 16.5 ± 0.5%, and 52.0 ± 0.6%, respectively, whereas 50% (w/w) Me-YCL resulted in 9.1 ± 1.3%, 30.7 ± 0.5%, and 67.8 ± 0.0%, respectively.Wound contraction plays a key role during the healing process as it decreases the wound size and promotes re-epithelization by minimising the distance traveled by migrating keratinocytes [56].Te increased rate of wound contraction in the animals treated with 50% (w/w) Me-YCL ointment may be attributed to its phenolic compounds which have antibacterial, anti-infammatory, and antioxidant properties [57].Additionally, the antioxidative properties of the leaf extract may have also contributed to the reduction of oxidative stress and subsequently led to the improvement of collagen synthesis, migration of myofbroblast, and proliferation of epithelial cells [58,59].Furthermore, by Day 12 and Day 14, percentage wound closure of more than 90% was observed in all the experimental groups, indicating the efcacy of Me-YCL and Me-MCL extracts in wound repair.
Te physical observation of the wound-healing progression for all the groups on Day 1, 3, 5, 7, 10, 12, and 14 can be seen in Table 4. Scab formation began to occur on Day 3 for all experimental groups, which may indicate the start of hemostasis, where platelet clots are formed to prevent further bleeding and protect from external contamination [60].In this phase, fbrin matrix is also generated which strengthens the initial seal and provides a framework for tissue restoration.Subsequently, detachment of these scabs can be seen on Day 10 for all treatment groups, while the scabs in the control groups were retained until Day 12.In this stage, angiogenesis is occurring, where formation of new blood vessel capillaries and granulation tissue is produced [61].Terefore, for the control groups with the scabs retained for a longer time, proliferation of new epithelial cells was delayed, and hence the rate of wound healing was reduced [58].
Te presence of bioactive compounds such as tannins, saponins, terpenoids, and glycosides in C. latifolia leaf extracts may have played a key role in the healing process.Teir overall bioactivities such as antimicrobial, antiinfammatory, and antioxidant could have provided a signifcant contribution to the rate of wound repair.For example, due to the antimicrobial and anti-infammatory activities in tannins and saponins, they can efectively Te Scientifc World Journal scavenge free radicals and enhance re-epithelialization and matrix synthesis of the wound area, restricting infammation that can delay the wound-healing process [62,63].As for glycosides and terpenoids, it has been reported that these compounds can promote collagen synthesis, along with fbroblast migration at the early proliferation phase of the wound-healing process [64,65].Tis is supported by the study of another species from the same genus known as Curculigo orchioides which showed that the wound-healing activity was improved with the facilitation of the phenolic glycoside compounds from the methanolic root extract such as corchioside A, issocrassifoside G, curculigosaponin A-F, sitosterol, linoleic acid, palmitic acid, and glucoside [63].Tese compounds were suggested to have enhanced the blood supply for vasoconstriction, epidermal cell migration, and re-arrangement of collagen fbers, thus stimulating the wound-healing activity [63].
Our preliminary study revealed a potential in woundhealing activity of methanolic young and matured C. latifolia leaf extracts on male Wistar rats.However, some modifcations to the study, such as increasing the dosage of extracts and exploring various formulations, can provide further improvements to the wound-healing process.Additional research is also needed to examine the impact of other extracts, such as ethanolic and aqueous extracts.Furthermore, an investigation into the diferent plant parts of C. latifolia, for example, the roots and rhizome, are essential for a comprehensive understanding of the plant's potential efects.

Conclusions
Te present study showed promising phytochemical, antioxidant, and antidiabetic activities in C. latifolia leaves and C. latifolia roots of Brunei Darussalam.Overall, C. latifolia roots presented the most prominent antioxidant and antidiabetic activities due to its phenolic content.Furthermore, ethanolic C. latifolia leaves have promising antihyperglycemic properties that could be greatly useful in the treatment of type 2 diabetes.Our study also demonstrated potential wound-healing activity by treatment with methanolic young and matured C. latifolia leaf extracts.However, a signifcant diference in comparison with control untreated group was only observed on Day 3, 5, and 7, and thus modifying the concentration levels of extract can potentially enhance the results.All the extracts studied have demonstrated some degree of bioactivity which could be utilised in the development of new drugs or health supplements for the treatment of various diseases.However, more studies such as their molecular pathways, compound isolation, and structure-activity relationship are needed to understand the mechanism of actions between the active components and their bioactivities.

Figure 4 :
Figure 4: Layout of measuring the wound contraction of male Wistar rat.

Figure 5 :
Figure 5: Average extraction yield of (a) sequential extracts and (b) ethanolic C. latifolia leaf (CL) and C. latifolia root (CR) extracts.Data are expressed as mean ± standard deviation (n � 3); * * p < 0.01 indicates the signifcant diference between solvents of low polarity and high polarity.
) and 7(d).Tis indicates that favonoids have little efect on the antioxidant activity and other major phenolic compounds in the ethanolic extracts may play a major role instead.

3. 4 .
Antidiabetic Activity by Chemical Assays.Preliminary screening of ethanolic C. latifolia leaf and C. latifolia root extracts for ∝ -glucosidase inhibitory activity and ∝ -amylase enzyme activity was carried out at 200 mg•L −1 , and the results of the study are shown in Figure 8.

Figure 6 :Figure 7 :
Figure 6: (a) TPC of sequential extracts at 500 mg/L, (b) TPC of ethanolic CL and CR extracts at 1000 mg/L, (c) TFC of sequential extracts at 500 mg/L, and (d) TFC of ethanolic CL and CR extracts at 1000 mg/L.Te statistical analysis showed signifcant diferences ( * * p < 0.01) between the extracts.

4. 1 .Figure 9 :
Figure 9: Blood glucose level in mmol/L of normal untreated control, diabetic control, and Curculigo latifolia leaf (CL) extract treatment group measured every week for a period of 14 weeks.* p < 0.05 indicates signifcant diferences between the experimental groups.Error bars denote standard deviation between BGL readings of animals (n � 6).

Figure 10 :
Figure 10: Blood glucose level in mmol/L portraying glucose tolerance of the animals in normal control, diabetic control, and CL treatment groups at 0, 15, 30, 60, 90, and 120 minutes.* p < 0.05 indicates signifcant diferences in the blood glucose levels between CL-treated group and alloxan diabetic control group at 120 min.Error bars denote standard deviation between BGL readings of animals (n � 6).
Chemical Assays 3.2.1.Qualitative Phytochemical Screening.Phytochemical screening for sequential extracts of C. latifolia young and matured leaves was carried out (results in Table 1(a)), which revealed the presence of terpenoids and glycosides, but no alkaloids in the extracts.Saponins and tannins were also present in all the extracts, except for the hexane and dichloromethane extracts.Similarly, results for ethanolic C. latifolia leaves of mixed-age and C. latifolia root extracts also showed presence of all the phytochemicals, except for alkaloids (see Table 1(b)).

Table 2 :
Te statistical correlation analysis of antioxidant activities with TPC and TFC of YCL and MCL and CL and CR extracts.
R value closer to 1 indicates a stronger correlation.

Table 3 :
Percentage change in body weight of normal control and diabetic control and CL treatment group at week 14 of the experimental period.
Statistical data marked signifcant diferences (p < 0.05) in percentage body weight change between experimental groups.Data are represented as mean ± standard deviation between body weights of animals (n � 6).